Cosmetic composition comprising white rose flower extract for skin whitening and improving skin wrinkle

ABSTRACT

Provided is a functional cosmetic composition of white rose extract and a gartanin derivative compound isolated therefrom, and specifically, to a cosmetic composition for skin whitening and skin wrinkle alleviation containing, as active ingredients, white rose petal extract and gartanin derivative compounds isolated therefrom. The white rose extract and gartanin derivative compound, according to the present invention, are safe without causing side effects on the skin, prevent melanin production through a mechanism inhibiting tyrosinase activity, thereby having a whitening effect, and exhibit a wrinkle alleviation effect by a mechanism inhibiting MMP-1 activity, and thus the composition, of the present invention, containing the same as active ingredients, can be utilized as a material for functional cosmetics for skin whitening and wrinkle alleviation without causing skin irritation.

TECHNICAL FIELD

The present invention relates to a functional cosmetic compositionincluding, as an active ingredient, white rose extract, and moreparticularly, to a functional cosmetic composition having skin whiteningand wrinkle alleviation functions including, as an active ingredient,white rose extract.

BACKGROUND ART

The skin is a defense organ on the front line of the human body whichprotects the body from external stimuli and even from the aestheticpoint of view, an interest thereof has been gradually increased.Accordingly, studies on a variety of functional cosmetics are activelyconducted and among the studies, various functional cosmetics forwrinkle alleviation and whitening functions are being launched.

Skin aging is largely divided into two types of intrinsic aging andextrinsic aging according to a cause thereof. The intrinsic aging is anaging process which is naturally caused while a physiological functionof the living body is deteriorated as the age increases and theextrinsic aging is an aging process generated by external factors suchas ultraviolet (UV), dry air, reactive oxygen species, and stress. Whenthe skin aging is in progress by these factors, wrinkle is generated onthe skin surface.

Various causes of the wrinkle generation have been reported and first,when the skin is exposed to excessive UV, decomposition of collagen as askin ingredient is accelerated and thus the skin elasticity is lost andthe wrinkle may be generated. Further, when the skin is excessivelydried by an excessive change in temperature, a decrease in humidity,wind, or the like, the skin function as a barrier to the outside isdeteriorated and the wrinkle is generated. In addition, when the skin isexposed to reactive oxygen species or free radicals, lipid peroxides areproduced by oxidation and as a result, skin structural proteins such ascollagen are deformed and the wrinkle may be generated.

Meanwhile, when the skin aging occurs, cornification of the skin occurs,a fusion rate of epidermal cells is gradually decreased, an epidermalcell layer is thinned, the cornified layer is thickened, and a boundarywith the dermal layer becomes in a straight line. In the corium, ashuman skin fibroblasts are aged, productivity of fibers and the matrixis reduced, intracellular peroxide levels rapidly increase, the functionof the cells cannot be smoothly performed and the metabolic activity ofthe cells is reduced under the influence of various types of pollutantsand stress. As a result, the wrinkle is generated due to functional andstructural damages and thus the skin aging is accelerated. Accordingly,various cosmetic compositions to prevent skin aging or wrinklegeneration caused by the external factor and the internal factors havebeen studied.

Meanwhile, when the UV is irradiated to the skin, melanocytes in theskin are activated and thus melanin production is accelerated by actionof enzyme tyrosinase, TRP1, and TRP2. The produced melanin is pigmentedin the skin to be developed to “age spots” and “freckles” and in orderto prevent the age spots and the freckles, whitening cosmetics includingvarious cosmetic compositions have been used. Further, it is known thatthe UV accelerates sebum, oxidation of a cell membrane and the like tocause various skin disorders, and particularly, recently, moreeffectively preventive measures of skin oxidation caused by an increasein UV according to depletion of the ozone layer have been required.

Ingredients having a whitening effect which are known in the related artinclude ascorbic phosphate ester salts, hydroquinone derivatives,placenta extracts, kojic acid, ellagic acid, and the like, and cosmeticcompositions mixing these ingredients are general. Particularly, inrecent years, whitening cosmetics using the placenta extracts have beenin the limelight, but in the placenta extracts, the supply amount islimited and the use there is limited in a combination of ethical issues.Accordingly, an interest in the development of novel and effectivewhitening ingredients has been focused.

In recent years, it is reported that polyphenol and the like included inthe plant have a high whitening effect and cosmetic compositions usingthe polyphenol and the like have been proposed. Further, the whiteningeffect of flavanones and hydroxy flavones is already disclosed. However,in these whitening ingredients, the effect thereof is somewhat minimaland there is a problem during storage, and a wrinkle prevention effectand an aging prevention effect in addition to the whitening effect arenot sufficiently exhibited. Particularly, there is a problem in thatfunctional cosmetics including chemical ingredients cause skinirritation and are not suitable for sensitive skin, and an interest infunctional cosmetics having small stimuli to the skin and derived fromeco-friendly natural plants has been focused.

Meanwhile, a white rose flower includes, as an active ingredient, awhite rose extract in the related art as a species belonging to thegenus Rose, and effects such as anti-oxidation, anti-inflammatory, andanti-allergy are already disclosed, but skin whitening and wrinklealleviation effects on the white rose flower are not yet studied.

Therefore, the inventors tested functions by focusing on a white roseextract while seeking a natural substance as a novel material forpreparing cosmetics with excellent skin whitening and wrinklealleviation effects without side effects to the living body. As aresult, the inventors found that an ethanol extract of the white roseeffectively inhibited activity of tyrosinase in vitro to have awhitening effect and effectively inhibited activity of collagenase tohave a wrinkle alleviation function and completed the present invention.

DISCLOSURE Technical Problem

The present invention is directed to provide a functional cosmeticcomposition having skin whitening and skin wrinkle alleviation effectswithout having side effects in the body.

Technical Solution

One aspect of the present invention provides a cosmetic composition forskin whitening or skin wrinkle alleviation including a white roseextract as an active ingredient.

The white rose extract may be an extract obtained by extracting driedwhite rose petal with ethanol, butanol, or ethyl acetate.

The white rose extract may prevent melanin production by suppressing orinhibiting activity of tyrosinase.

The white rose extract may improve skin elasticity by suppressing orinhibiting activity of MMP-1.

Another aspect of the present invention provides a cosmetic compositionfor skin whitening or skin wrinkle alleviation including3,5-di-o-methyl-gartanin as an active ingredient.

The 3,5-di-o-methyl-gartanin may be isolated from at least one roseselected from the group consisting of white rose flower, Red Sandra, RedVelvet, First Red, Nobless, Konffetti, My Heart, Grand Gala, Dolores,Rote Rose, Saphir, Mercedes, Gabriella, Frisco, Only Love, Coco, Escimo,Calibra, Mimi Rose, Little Mable, Princess, Evelien and Chaming. The3,5-di-o-methyl-gartanin may be derived from the white rose petalextract.

The 3,5-di-o-methyl-gartanin may prevent melanin production bysuppressing or inhibiting activity of tyrosinase.

The 3,5-di-o-methyl-gartanin may improve skin elasticity by suppressingor inhibiting activity of MMP-1.

The composition may be formulated by at least one selected from thegroup consisting of skin lotion, skin softener, skin toner, astringent,lotion, milk lotion, moisturizing lotion, nutrition lotion, massagecream, nourishing cream, moisturizing cream, hand cream, essence,nutrition essence, pack, soap, shampoo, cleansing foam, cleansinglotions, cleansing cream, body lotion, body cleanser, emulsion,lipstick, makeup base, foundation, pressed powder and loose powder.

Advantageous Effects

3,5-di-o-methyl-gartanin derived from a white rose petal extract orwhite rose petal has a whitening effect by suppressing melaninproduction through a mechanism of inhibiting tyrosinase activity and haseffects of enhancing skin elasticity and alleviating a wrinkle through amechanism of inhibiting activity of MMP-1. Therefore, the composition ofthe present invention including 3,5-di-o-methyl-gartanin as an activeingredient can be utilized as a material for functional cosmetics forskin whitening and skin wrinkle alleviation without causing skinirritation.

DESCRIPTION OF DRAWINGS

FIG. 1 is a result of observing an MMP-1 inhibitory effect for anextract obtained by extracting white rose petal with ethanol, butanol,and ethyl acetate (black: 1 μg/mL, red: 3.2 μg/mL, green: 10 μg/mL,yellow: 32 μg/mL, and blue: 100 μg/mL).

FIG. 2 is a result of observing a tyrosinase inhibitory effect for anextract obtained by extracting white rose petal with ethanol, butanol,and ethyl acetate (black: 1 μg/mL, red: 3.2 μg/mL, green: 10 μg/mL,yellow: 32 μg/mL, and blue: 100 μg/mL).

FIG. 3 is a result of observing a melanin synthesis inhibitory effectfor an extract obtained by extracting white rose petal with ethanol,butanol, and ethyl acetate (black: 1 μg/mL, red: 3.2 μg/mL, green: 10μg/mL, yellow: 32 μg/mL, and blue: 100 μg/mL).

FIG. 4 is a result illustrating a process of preparing white rose petalfractions of the present invention.

FIG. 5 is a result of measuring DPPH radical scavenging activity foreach treatment concentration with respect to extracts of the white rosepetal extracted with ethanol, butanol, and fraction 2.

FIG. 6 is a result of performing a metal ion catalytic oxidationinhibition test for each treatment concentration with respect toextracts of the white rose petal extracted with ethanol, butanol, andfraction 2.

FIG. 7 is a result of measuring tyrosinase activity for each treatmentconcentration with respect to extracts of the white rose petal extractedwith ethanol, butanol, and fraction 2.

MODES OF THE INVENTION

While the inventors studied a novel cosmetic material capable ofpreventing skin aging, alleviating skin wrinkle, and enhancing a skinwhitening effect, the inventors verified that a white rose petal extracthad the functions.

Therefore, the present invention is characterized by providing afunctional cosmetic composition including, an active ingredient, a whiterose petal extract extracted from white rose petal.

Particularly, while the inventors was trying to find a material havingan excellent whitening effect and anti-aging effect while being derivedfrom a natural material to be stable in the human body, the inventorspaid attention to a white rose extract and verified that the white rosepetal extract had a whitening effect and a wrinkle alleviation effectthrough an experiment.

That is, according to an exemplary embodiment of the present invention,it is exhibited that the white rose extract suppresses or inhibitsactivity of MMP-1 to improve skin elasticity (see FIG. 1).

Meanwhile, melanin as a pigment present in the skin has an importantfunction of protect a body from external stimuli such as UV rays.However, when the melanin is excessively produced to be pigmented, themelanin forms age spots, freckles, or the like to cause skin appearanceproblems, and furthermore, facilitate skin aging and cause skin cancer,and thus a material capable of suppressing over-production of themelanin can be used as a whitening product.

It is known that skin pigmentation is caused by stimuli such as UV orinflammation and generated by genetic factors or factors such ashormones, chemicals, foods, and the like. Skin melanocytes are involvedin a biosynthesis process of melanin. That is, the biosynthesis of themelanin occurs in a specific-shaped small organ in melanocytes calledmelanosome and an enzyme called tyrosinase plays the most important rolein generation of the melanin in the melanosome. Tyrosine is converted todopa by action of the enzyme, generated to dopaquinone through a seriesof oxidation processes, and converted to dopachrome again to finallyform dark-brown melanin. The expression of tyrosinase, tyrosinaserelated protein-1 (TRP-1), TRP-2, and extracellular signal-regulatedkinases (ERK) as protein genes involved in formation of the melaninpigment and a microphthalmia-associated transcription factor (MITF)which is a main transcription factor regulating expression of themelanin synthesis plays an important role in a melanin synthesisprocess.

According to an exemplary embodiment of the present invention, it isexhibited that the white rose petal extract of the present inventioninhibits tyrosinase concentration-dependent to have activity ofsuppressing melanin synthesis (see FIGS. 2 and 3).

Accordingly, the white rose petal extract of the present invention ofsuppressing the tyrosinase activity may be used for preventing skinpigmentation and a skin whitening effect.

Through such a result, the inventors were experimentally verified thatthe white rose petal extract of the present invention has both the skinwrinkle alleviation effect and the skin whitening effect.

Meanwhile, the inventors additionally experimented whether any componentof the white rose petal extract particularly has the skin wrinklealleviation effect and the skin whitening effect.

As a result, it was verified that 3,5-di-o-methyl-gartanin correspondingto a 14-th peak of a fraction obtained by extracting white rose petalwith butanol has the best MMP-1 activity inhibition and tyrosinaseactivity inhibition.

Accordingly, the present invention may provide a cosmetic compositionfor skin whitening and skin wrinkle alleviation including, as activeingredients, 3,5-di-o-methyl-gartanin in addition to the white rosepetal extract.

The white rose extract according to the present invention may use anextract extracted and isolated from a natural material by usingextraction and isolation methods which are known in the art. The“extract” defined in the present invention is extracted from white roseby using an appropriate solvent and for example, includes all of a crudeextract, a polar solvent soluble extract, and a non-polar solventsoluble extract of the white rose. Preferably, the white rose extractmay be an extract extracted from white rose petal.

An appropriate solvent for extracting the extract from the white rosemay use any pharmaceutically acceptable organic solvent and may usewater or an organic solvent, but is not limited thereto. For example,the solvent may use various solvents including purified water, alcoholsof carbon atoms 1 to 4 including methanol, ethanol, propanol,isopropanol, butanol, and the like, acetone, ether, benzene, chloroform,ethyl acetate, methylene chloride, hexane, cyclohexane, and the like,alone or in combination. Preferably, the solvent may use ethanol(alcohol), butanol, or ethyl acetate.

The extraction method may use any one selected from methods includinghot water extraction, chilling extraction, reflux cooling extraction,solvent extraction, steam distillation, ultrasonic extraction, elution,pressing, and the like. Further, a desired extract may additionallyperform a general fraction process and be purified by using a generalpurification method. The method of preparing the white rose extract ofthe present invention is not limited and may use any known method.

For example, in the white rose extract included in the composition ofthe present invention, a primary extract extracted by the hot waterextraction or solvent extraction may be prepared in a powder state by anadditional process such as vacuum distillation and freeze drying orspray drying. Further, the primary extract may obtain additionallypurified fractions by using various chromatography including silica gelcolumn chromatography, thin layer chromatography, high performanceliquid chromatography, and the like.

Accordingly, in the present invention, the white rose extract is aconcept including all of extracts, fractioned and purified materials,their dilutions, concentrates, or dried materials which are obtained ineach step of extraction, fraction, or purification.

The method of preparing the white rose extract according to theexemplary embodiment of the present invention will be described below inmore detail.

In the present invention, the dried white rose petal is grinded andpowdered and then extracted for 24 hours at room temperature by addingethanol or water to the powder. Thereafter, the water extract isadditionally deposited with butanol or ethyl acetate for 24 hours atroom temperature again to obtain the white rose petal extract through aprocess which is prepared to the freeze-dried powder.

Further, 100 mg of the butanol extract is centrifuged for 2 minutes at10,000 rpm after adding 1 ml of methanol and F2 fractioned by adding 1ml of butanol to the precipitated pellet is analyzed by using GC/MS(agilent technologies 5975C), and as a result, 3,5-di-o-methyl-gartaninmay be obtained through a 14-th solvent peak.

As such, when the extract extracted from the white rose petal or acompound purified therefrom is prepared by the cosmetic composition, thecomposition of the present invention may include ingredients generallyused in the cosmetic composition in addition to the aforementioned whiterose petal extract, for example, general additives, such asantioxidants, stabilizers, solubilizers, vitamins, pigments andperfumes, and carriers.

Further, the composition of the present invention may be used bycombining an organic sunscreen which has been used from the past to theextent that a skin protection effect is not damaged by reacting with thewhite rose petal extract other than the aforementioned white rose petalextract.

The organic sunscreen may use at least one selected from the groupconsisting of glyceryl PABA, drometrizole trisiloxane, drometrizole,digalloyl trioleate, disodium phenyl dibenzimidazole tetrasulfonate,diethylhexyl butamido triazone, diethylamino hydroxybenzoyl hexylbenzoate, DEA-methoxylcinnamate, a mixture of lawsone and dihydroxylacetone, methylene bis-benzotriazolyl tetramethylbutylphenol,4-methylbenzylidene camphor, menthyl anthranilate, benzophenone-3(oxybenzone), benzophenone-4, benzophenone-8 (dioxyphebenzone), butylmethoxydibenzoylmethane, bis-ethylhexyloxyphenolmeth oxyphenyltriazine,cinoxate, ethyldihydroxypropyl PABA, cctocrylene, ethylhexyl dimethylPABA, ethylhexyl methoxycinnamate, ethylhexyl salicylate, ethylhexyltriazone, isoamyl p-methoxycinnamate, polysilicon-15(dimethicodiethylbenzal malonate), terephthalylidene dicamphor sulfonicacid and its salts, TEA salicylate, and amino benzoic acid (PABA).

Further, the cosmetic composition of the present invention may beprepared by any formulation which is generally prepared in the art andfor example, may be formulated by a solution, a suspension, an emulsion,paste, gel, cream, lotion, powder, soap, a surfactant-containingcleanser, oil, powder foundation, emulsion foundation, wax foundation,spray and the like, but is not limited thereto. More particularly, thecosmetic composition of the present invention may be prepared byformulation of emulsion lotion, nutrition lotion, nourishing cream,massage cream, essence, eye cream, cleansing cream, cleansing foam,cleansing water, pack, spray or powder.

When the formulation of the present invention is paste, cream, or gel,as a carrier ingredient, animal oil, vegetable oil, waxes, paraffins,starch, tragacanth, cellulose derivatives, polyethylene glycols,silicones, bentonites, silica, talc, zinc oxide, or the like may beused.

When the formulation of the present invention is the solution or theemulsion, as a carrier ingredient, a solvent, a dissolving agent, or anemulsifying agent is used, and for example, water, ethanol, isopropanol,ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate,propylene glycol, 1,3-butyl glycol oil, glycerol aliphatic ester,polyethylene glycol, or sorbitan fatty acid ester is included.

When the formulation of the present invention is the suspension, as thecarrier ingredient, a liquid diluent, such as water, ethanol, orpropylene glycol, a suspension such as ethoxylated isostearyl alcohol,polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester,microcrystalline cellulose, aluminum metahydroxide, bentonite, agar ortragacanth may be used.

When the formulation of the present invention is the powder or thespray, as the carrier ingredient, lactose, talc, silica, aluminumhydroxide, calcium silicate, or polyamide powder may be used.Particularly, in the case of the spray, a propellant such aschloro-fluoro hydrocarbon, propane/butane or dimethyl ether may beadditionally included.

When the formulation of the present invention is thesurfactant-containing cleanser, as the carrier ingredient, aliphaticalcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinatemonoester, isethionate, imidazolinium derivatives, methyl taurate,sarcosinate, fatty acid amide ether sulfate, alkyl amido betaine,aliphatic alcohol, fatty acid glyceride, fatty acid diethanolamide,vegetable oil, lanoline derivatives, ethoxylated glycerol fatty acidester, or the like may be used.

Further, the present invention may provide a make-up method having askin aging prevention, skin wrinkle alleviation, or skin whiteningeffect by applying a cosmetic composition including, as an activeingredient, a white rose petal extract or 3,5-di-o-methyl-gartanin tothe human skin.

The make-up method of the present invention refers to all make-upmethods using the cosmetic composition of the present invention. Thatis, all of the methods which are known in the art using the cosmeticcomposition belong to the make-up method of the present invention.

The cosmetic composition of the present invention is used alone or induplicate or may be used by duplicating other cosmetic compositionsother than the cosmetic composition of the present invention. Further,the cosmetic composition according to the present invention may be usedaccording to a general use method and vary the number of uses accordingto a skin state or a taste of a user.

When the cosmetic composition of the present invention is soap,surfactant-containing cleansing formulation or a surfactant-freecleansing formulation, the cosmetic composition is coated on the skinand then wiped or removed or cleaned with water. As a particularexample, the soap is liquid soap, powder soap, solid soap and oil soap,the surfactant-containing cleansing formulation is cleansing foam,cleansing water, a cleansing towel and a cleansing pack, and thesurfactant-free cleansing formulation is cleansing cream, cleansinglotion, cleansing water and cleansing gel, and the present invention isnot limited thereto.

Hereinafter, the present invention will be described in more detail byExamples. These Examples are just to describe the present invention inmore detail and it is apparent to those skilled in the art that thescope of the present invention is not limited to these Examples.

Example 1 Preparation of White Rose Petal

The white rose petal used in the present invention was collected atJincheon, Chungbuk, Korea in May, 2010 and then completely dried underthe sun and used. The completely dried petal was grinded and powdered byusing a rotor speed mill (Laval Lab Inc., Laval, Que), sterilized byusing a 70% ethanol spray, dried at 80° C., and then stored at 4° C.before usage.

Example 2 Preparation of White Rose Petal Extract

The prepared dried white rose petal powder was divided into two partsand extracted with ethanol or water.

First, the ethanol extract was deposited by using 70% ethanol 10 timeslarger than a volume of the white rose petal powder and extracted for 24hours at room temperature. An extracted suspension was filtered by aWhattman filter (No. 1) and the filtrate was concentrated under a vacuumat 50° C. and freeze-dried to obtain the extract. Samples obtained fromthe ethanol extract were continuously fractioned through n-hexane ethylacetat (EA), chloroform, n-butanol (BuOH), and distilled water, and theobtained fractions were dried by an evaporator. Then, the obtainedproducts were used for the experiments.

Example 3 Analysis of Collagen Matrix Metalloproteinase-1 InhibitoryEffect of White Rose Petal Extract

As part of measuring a wrinkle alleviation effect of the white rosepetal extract, effects of inhibiting activity of collagen matrixmetalloproteinase-1 (MMP-1) of the extracts were measured with referenceto the guideline of “beauty-related functional tests (pp. 821-858)” ofKorea health official compendium research society foundation.

When simply describing the effect, 25 mg of bovine tendon collagen wasdissolved in a 0.05 M tris(hydroxymethyl)-methyl-2-aminoethane sulfonate(TES) buffer and then left for 15 minutes at 37° C. Next, a testmaterial for each concentration was added and then additionally reactedwith collagenase type-1 for 5 hours at 37° C. 0.2 mL of a reactionsolution was taken and added to 1 mL of a buffer [14% ninhydrin inmethyl cellosolve (2-methoxyethanol)+0.2 M sodium citrate with 0.71 mMstannous chloride]. The reaction solution was boiled for 20 minutes,added with n-propanol, and then left for 15 minutes and then absorbancewas measured at 600 nm. On the basis of a change in absorbance when onlythe buffer without the white rose fractions was added as a reference(100%), reduction of the absorbance caused by the white rose fractionsfor each concentration was represented by an inhibition rate.

As a result, as illustrated in FIG. 1, the ethanol fraction exhibited aMMP-1 activity inhibition rate of 41.7% in 100 μg/mL, and the butanolfraction exhibited the MMP-1 activity inhibition rate of 34.9% in 10μg/mL, 47.7% in 32 μg/mL, and 76.1% in 100 μg/mL. Meanwhile, the ethylacetate fraction had the strongest MMP-1 activity inhibition effect andexhibited the inhibition rate of 35.0% in 3.2 μg/mL, 39.8% in 10 μg/mL,59.0% in 32 μg/mL, and 83.1% in 100 μg/mL.

Through the above result, the inventors verified that all of the ethanolextract, the butanol extract, and the ethyl acetate extract of the whiterose petal inhibited the activity of MMP-1. Accordingly, it can be seenthat the ethanol extract, the butanol extract, and the ethyl acetateextract of the white rose petal have an excellent wrinkle alleviationeffect.

Example 4 Analysis of Melanin Synthase (Tyrosinase) Inhibitory Effect ofWhite Rose Petal Extract

Meanwhile, in order to measure a skin whitening effect of the white rosepetal extract, an effect of inhibiting activity of tyrosinase as melaninsynthase was measured with reference to the guideline of “beauty-relatedfunctional tests (pp. 821-858)” of Korea health official compendiumresearch society foundation.

When simply describing this, 20 μL of mushroom tyrosinase (2,000unit/mg) and 10 μL of 10 mM L-dihydroxyphenylalanine (L-DOPA) were addedin 70 μL of a white rose fraction sample (1, 3.2, 10, 32, 100 μg/mL offinal concentrations) dissolved in PBS (pH 6.8) on each well of a96-well plate and then reacted for 30 minutes at 37° C., and absorbanceat 475 nm was measured by using a microplate reader (Infinite 200, TecanGroup Ltd., Switzerland). In this case, on the basis of a change inabsorbance when only the buffer without the white rose fractions wasadded as a reference (100%), reduction of the absorbance caused by thewhite rose fractions for each concentration was represented by aninhibition rate.

As a result, as illustrated in FIG. 2, the ethanol fraction exhibited atyrosinase inhibition rate of 29.4% in 32 μg/mL and 52.9% in 100 μg/mL,and the butanol fraction exhibited the tyrosinase inhibition rate of38.1% in 32 μg/mL, and 60.4% in 100 μg/mL. Meanwhile, it was verifiedthat the ethyl acetate fraction exhibited the strongest tyrosinaseinhibition rate and exhibited a tyrosinase inhibition rate of 44.3% in32 μg/mL and 65.1% in 100 μg/mL.

Example 5 Analysis of Melanin Synthesis Inhibitory Effect of White RosePetal Extract

In order to more definitize a skin whitening effect of the white rosepetal extract, the inventors measured a melanin synthesis inhibitoryeffect of the white rose petal extract at a cell level with reference tothe guideline of “beauty-related functional tests (pp. 821-858)” ofKorea health official compendium research society foundation (2004).

When simply describing this, B16 mouse melanoma cells were dispensed to1×10⁴ cells/well (0.5 mL) in a 96-well plate and cultured for 24 hours.The B16 mouse melanoma cells was treated with the white rose fractionfor each concentration and a α-melanocyte stimulating hormone (MSH) andadditionally cultured for 3 days at 37° C. The cells were lysed byadding 0.15 mL of 1 M NaOH to a culture medium and boiled for 5 minutesto elute the melanin. 0.1 mL of the eluate was taken and absorbance at490 nm was measured. A standard calibration curve was created bysynthetic melanin and the concentration was quantified and an inhibitionrate for each concentration of the white rose fraction was proposedbased on a melanin production amount when the sample was not treated asa reference (100%).

As a result, as illustrated in FIG. 3, the ethanol fraction exhibited amelanin synthesis inhibition rate of 27.7% in 32 μg/mL and 39.9% in 100μg/mL, and the butanol fraction exhibited the melanin synthesisinhibition rate of 23.5% in 10 μg/mL, 35.5% in 32 μg/mL, and 49.6% in100 μg/mL. Meanwhile, the ethyl acetate fraction exhibited the strongestmelanin synthesis inhibition rate and exhibited the melanin synthesisinhibition rate of 30.2% in 10 μg/mL, 54.4% in 32 μg/mL, and 58.8% in100 μg/mL.

Through the result, the inventors verified that all of the ethanolextract, the butanol extract, and the ethyl acetate extract of the whiterose petal inhibited the activity of the tyrosinase and suppressed themelanin synthesis. Accordingly, it can be seen that he ethanol extract,the butanol extract, and the ethyl acetate extract of the white rosepetal exhibit the excellent whitening effect.

Example 6 Ingredient Analysis of White Rose Petal Extract

The inventors analyzed specific ingredients of the white rose petalextract having the wrinkle alleviation and whitening effects. To thisend, the inventors isolated a supernatant (fraction 1, 75%, hereinafter,referred to as ‘F1’) by adding 1 ml of methanol to 100 mg of the butanolextract and then the centrifuging the butanol extract for 2 minutes at10000 rpm and obtained fraction 2 (8%, hereinafter, referred to as ‘F2’)by adding 1 ml of butanol to the precipitated pellet (See FIG. 4).

The F1 and F2 were analyzed by using GC/MS (Agilent Technologies 5975C)installed with a CTC CombiPAL autosampler system (Palo Alto, Calif.,USA). A column used in sample analysis was a HP-5 column (AgilentTechnologies, 250 μm×0.25 μm×30 m) and 10 pt of the sample was analyzed.An injector temperature was 250° C. and was measured up to 280° C. at aninterval of 3° C. per minute after a GC oven stopped for 5 minutes at50° C. Data from after 4 minutes was collected by considering a solventpeak and Wiley7N library data were used for ingredient analysis.

As a result, ingredients in the following Tables 1 and 2 could beverified.

TABLE 1 Ingredient Analysis Table of F1 Retention Peak time Fraction 1 %Area Quality 1 6.448 2-Methyl-2-butanal 0.37 33 2 7.518 Furfural 5.60 643 7.701 1,5-Dimethyl-1H-pyrazole 0.13 86 4 8.192 2,5-Furandione 0.19 865 12.249 4-Oxo-5-methoxy-2-penten-5- 0.27 86 olide 6 12.5005-methyl-2-furancarboxaldehyde 0.38 95 7 13.1651-Methoxy-difluorobenzene 0.20 38 8 13.8783-Acetyldihydro-2(3H)-furanone 0.18 50 9 14.572 Propynamide 0.16 37 1016.113 3-Fluoro-1,2-benzenediol 0.40 27 11 16.219 4-Ethyl-cyclohexanone0.12 30 12 16.297 4-Oxo-pentanoic acid 0.37 14 13 17.6462-Methoxy-6-methyl-pyrazine 0.10 74 14 18.1473-Hydroxy-5-methyl-4H-pyran-4- 0.18 35 one 15 19.322 2-Furanmethanol0.26 86 16 20.084 5-Methyl-2(3H)-furanone 0.12 30 17 21.0282,3-Dihydro-3,5-dihydroxy-6- 0.89 90 methyl-4H-pyran-4-one 18 23.6302-Butyl-4,5-dimethyloxazole 0.24 38 19 23.909 Butanedioic acid 0.43 2720 24.256 3,4,5,6-Dibenzocarbazole 2.29 50 21 24.6612-Methyl-cycloheptanone 0.45 30 22 24.941 5-(hydroxymethyl)-2- 31.56 91Furancarboxaldehyde 23 29.624 1-(3-Chloro-2-propenyl)-4- 0.58 30 methoxybenzene 24 29.778 1-Fluoro-3(2-cyanoethenyl)- 0.62 22 benzene 25 29.9714-Methoxy-benzeneacetonitrile 1.05 46 26 30.000 1a,2,3,5-tetrahydro-1H-1.28 47 cycloprop[c]indol-5-one 27 30.858 1,3-cyclopentadione 0.53 30 2831.330 1,2,3-Benzenetriol 40.38 95 29 40.195 Harmin 0.17 47 30 40.2341,2-Dihydro-7,12-methano-4H- 0.11 64 cyclodeca[c]pyran-4-one 31 40.330Tetrahydroanthraquinone 0.09 64 32 40.542 (E)-2,4′-dihydroxystilbene0.66 64 33 40.610 [2.2](2,6)pyrazinophane 0.24 83 (E)-2,2′-stilbenediol50 34 40.976 1-(4- 3.49 43 Pyrroylcarbonyl)benzotriazole 35 41.988(Z)-1,2-Difluoro-1-(p- 0.58 50 methoxyphenyl)ethene 36 75.8613,5-Di(p-chlorophenyl)-isoxazole 0.21 43

TABLE 2 Ingredient Analysis Table of F2 Retention Peak time Fraction 2 %Area Qality 1 6.217 n-Butyl acetate 3.15 72 2 9.252 4-Heptanone 1.42 863 9.464 n-Butyl ether 4.12 90 4 11.257 3-Methyl-4-heptanone 1.27 95 514.08 Butyl butylate 2.46 90 6 19.303 5-phenyl-8-oxa-1- 1.17 59azabicyclo[3.2.1]octane 3-phenyl-1,2,4-triazine- 59 5,6(1H,2H)-dione 723.331 1-Methyl-azetidine 0.89 25 8 25.827 1,3-bis(1,1-dimethylethyl)-0.74 9 benzene 9 32.004 Dihexylsulfide 0.66 43 10 33.451,1′-(1,4-phenylene)bis-ethanone 0.84 86 1-(1,1-dimethylethyl)-3,5- 80dimethyl-benzene 11 35.175 Iso butanol 0.52 5 12 72.189 Docosane 0.87 3513 73.008 Heptacosane 1.12 14 14 76.738 3,5-di-o-methyl-gartanin 1.35 35

Example 7 Analysis of Collagen Matrix Metalloproteinase-1 InhibitoryEffect for Each Ingredient of White Rose Petal Extract

The inventors tested the MMP-1 inhibitory effect of 36 ingredientsanalyzed in the F1 and 14 ingredients analyzed in the F2 in order toverify which ingredient of the white rose petal extract had the skinwrinkle alleviation effect.

The test method was performed the same as the method disclosed inExample 3 and as the tested result, it was verified that in3,5-di-o-methyl-gartanin which was the 14-th fraction of the F2, aparticularly excellent inhibition effect of MMP-1 was exhibited.

Example 8 Analysis of Melanin Synthase (Tyrosinase) Inhibitory Effect of3,5-Di-o-Methyl-Gartanin

The inventors performed the test by the same method as the methoddisclosed in Example 4 in order to verify whether3,5-di-o-methyl-gartanin showing the excellent MMP-1 inhibitory effectexhibited the tyrosinase inhibitory effect as the melanin synthase.

As a result, it was verified that 3,5-di-o-methyl-gartanin had anexcellent tyrosinase inhibition activity.

Example 9 Analysis of Melanin Synthesis Inhibitory Effect of3,5-Di-o-Methyl-Gartanin

Finally, the inventors observed the melanin synthesis inhibitory effectdue to the tyrosinase inhibition activity of the3,5-di-o-methyl-gartanin through the test method disclosed in Example 5.

As a result, expectedly, the inventors verify that the melanin synthesisof the B16 mouse melanoma cells may be efficiently inhibited bytreatment of 3,5-di-o-methyl-gartanin.

Example 10 Analysis of DPPH Reactive Oxygen Scavenging Activity of WhiteRose Petal Extract for Each Treatment Concentration

2,2-Diphenyl-1-picrylhydrazyl (DPPH) is a free radical which is verystable by itself and a dark violet compound having characteristic lightabsorption at 517 nm, but is quantitatively discolored by antioxidantshaving radical scavenging activity. Further, the radical scavengingactivity has a correlation with antioxidation activity includinginhibition activity of lipid peroxidation and is an experimental methodwhich is widely used for searching antioxidants.

Accordingly, in order to verify DPPH reactive oxygen scavenging activityof the white rose flower, the inventors detected reaction of a DPPHsolution, ethanol (EtOH) extract, and each fraction and verified theDPPH reactive oxygen scavenging activity by measuring absorbance at 517nm by using a microplate-ELISA reader (SpectraMax plus 384, MolecularDevices, CA, USA) after treating vitamin C (Vit.C) as a positive controlgroup.

The DPPH reactive oxygen scavenging activity was measured after treatingwhite rose flower samples with concentrations of 1, 10, 25, and 50 μg/mLrespectively, and as a result, it can be seen that the DPPH reactiveoxygen scavenging activity is exhibited from the concentration of 10μg/mL in all the samples (see FIG. 5).

Example 11 Analysis of Metal Ion Catalytic Oxidation Inhibition of WhiteRose Petal Extract for Each Treatment Concentration

As a method of verifying an effect of antioxidants protecting proteinssuch as bovine serum albumin (BSA) or enzymes from the damage byreactive oxygen species (ROS) by using a metal ion catalytic reaction,in order to verify metal ion catalytic oxidation inhibition activity ofthe white rose petal extract and the fractions, the following experimentwas performed.

First, in order to induce production of free radicals, the metal ioncatalytic reaction was used by adding CuSO₄ 1 mM and H₂O₂ 10 mM and freeradicals were produced and then electrophoresed in SDS-polyacrylamidegel for a protection effect of the BSA protein of the extracts and thefractions.

A protein protection effect to the metal ion catalytic free radicalsafter treating the white rose flower samples with concentrations of 1,10, 25, and 50 μg/mL was evaluated. As a result, it can be seen that inan ethanol (EtOH) extract, a protein protection effect by the metal ioncatalytic free radicals is exhibited from the concentration of 10 μg/mL,whereas in butanol (BuOH) and Fraction 2, the protection effect isobserved from the concentration of 1 μg/mL (see FIG. 6).

Example 12 Analysis of Tyrosinase Activity Inhibition In Vitro of WhiteRose Petal Extract for Each Treatment Concentration

The inventors mixed the white rose flower fractions with 100 mM of asodium phosphate buffer (pH 6.5), L-tyrosine, and mushroom tyrosinaseand reacted at 37° C. for 30 minutes, and thereafter, measuredtyrosinase inhibition activity through intensity after photographing byusing a OLYMPUS CKX 41 (Olympus, Tokyo, Japan).

The tyrosinase inhibition activity after treating the white rose flowersamples with concentrations of 10 and 25 μg/mL was measured. As aresult, it can be seen that the tyrosinase inhibition activity isexhibited in all the samples and particularly, in butanol (BuOH) andFraction 2, the tyrosinase inhibition activity is exhibited at a similarlevel to vitamin C as a positive control group (see FIG. 7).

Through the above result, the inventors experimentally verified that thewhite rose petal extract had the skin whitening and skin wrinklealleviation effects and verified that the white rose petal extract,particularly, the 3,5-di-o-methyl-gartanin exhibited an excellent effectin skin whitening and skin wrinkle alleviation by the MMP-1 activity andtyrosinase activity inhibition effects. Accordingly, the cosmeticcomposition including the white rose petal extract or the3,5-di-o-methyl-gartanin can be utilized as an important ingredient forfunctional cosmetics for skin whitening and skin wrinkle alleviation.

Hereinafter, as cosmetic Formulation Examples of the present invention,Lotion, essence, body cleanser, and the like including a white roseextract (alternatively, 3,5-di-o-methyl-gartanin) as an activeingredient of the present invention will be exemplified. However, theformulation of the cosmetic material of the present invention is notnecessarily limited thereto.

Formulation Example 1 Skin Lotion

A skin lotion was prepared by a general method by adding the followingingredients and contents.

White rose extract (alternatively, 3,5-di-o-methyl-gartanin)—2.0 wt %,1,3-butylene glycol—5.2 wt %, oleyl alcohol—1.5 wt %, ethanol—3.2 wt %,polysorbate 20-3.2 wt %, benzophenone 9-2.0 wt %, carboxyl vinylpolymer—1.0 wt %, glycerine—3.5 wt %, flavor—small amount,preservative—small amount, and purified water—residual amount %

Formulation Example 2 Essence

An essence was prepared by a general method by adding the followingingredients and contents.

White rose extract (alternatively, 3,5-di-o-methyl-gartanin)—2.0 wt %,cetostearyl alcohol—1.0 wt %, glyceryl monostearate—0.8 wt %, sorbitanmonostearate—0.3 wt %, propylparaben—0.1 wt %, polysorbate 60-1.0 wt %,mineral oil—5.0 wt %, cyclomethicone—3.0 wt %, dimethicone—0.5 wt %,allantoin—0.1 wt %, glycerine—5.0 wt %, alcohol—2 wt %, propyleneglycol—3.0 wt %, flavor—small amount, preservative—small amount, andpurified water—residual amount %

Formulation Example 3 Body Cleanser

A body cleanser was prepared by a general method by adding the followingingredients and contents.

White rose extract (alternatively, 3,5-di-o-methyl-gartanin)—2.0 wt %,alkylether saccharate salt (30%)—30.0 wt %, alkylether sulfosuccinicacid salt (30%)—20.0 wt %, alkylamido betaine (30%)—5.0 wt %, propyleneglycol—5.0 wt %, betaine—5.0 wt %, lauramide DEA—4.0 wt %, cellulosegum—0.05 wt %, hydrolyzed protein—0.1 wt %, citric acid—small amount,NaCl—small amount, flavor—small amount, preservative—small amount, andpurified water—residual amount %

Hereinabove, the preferred exemplary embodiments of the presentinvention have been primarily described. It is understood to thoseskilled in the art that the present invention may be implemented as amodified form without departing from an essential characteristic of thepresent invention. Therefore, the disclosed exemplary embodiments shouldbe considered from not a limitative viewpoint but an explanatoryviewpoint. It should be construed that the scope of the presentinvention should be interpreted by not the above description but theappended claims and all differences in the equivalent range thereto areincluded in the present invention.

1. A cosmetic composition for skin whitening or skin wrinklealleviation, the cosmetic composition comprising a white rose extract asan active ingredient.
 2. The cosmetic composition of claim 1, whereinthe white rose extract is an extract obtained by extracting dried whiterose petal with ethanol, butanol, or ethyl acetate.
 3. The cosmeticcomposition of claim 1, wherein the white rose extract prevents melaninproduction by suppressing or inhibiting activity of tyrosinase.
 4. Thecosmetic composition of claim 1, wherein the white rose extract improvesskin elasticity by suppressing or inhibiting activity of MMP-1.
 5. Acosmetic composition for skin whitening or skin wrinkle alleviation, thecosmetic composition comprising 3,5-di-o-methyl-gartanin as an activeingredient.
 6. The cosmetic composition of claim 5, wherein the3,5-di-o-methyl-gartanin is isolated from at least one rose selectedfrom the group consisting of white rose flower, Red Sandra, Red Velvet,First Red, Nobless, Konffetti, My Heart, Grand Gala, Dolores, Rote Rose,Saphir, Mercedes, Gabriella, Frisco, Only Love, Coco, Escimo, Calibra,Mimi Rose, Little Mable, Princess, Evelien and Chaming.
 7. The cosmeticcomposition of claim 6, wherein the 3,5-di-o-methyl-gartanin is derivedfrom the white rose petal extract.
 8. The cosmetic composition of claim5, wherein the 3,5-di-o-methyl-gartanin prevents melanin production bysuppressing or inhibiting activity of tyrosinase.
 9. The cosmeticcomposition of claim 5, wherein the 3,5-di-o-methyl-gartanin improvesskin elasticity by suppressing or inhibiting activity of MMP-1.
 10. Thecosmetic composition of claim 5, wherein the composition is formulatedby at least one selected from the group consisting of skin lotion, skinsoftener, skin toner, astringent, lotion, milk lotion, moisturizinglotion, nutrition lotion, massage cream, nourishing cream, moisturizingcream, hand cream, essence, nutrition essence, pack, soap, shampoo,cleansing foam, cleansing lotions, cleansing cream, body lotion, bodycleanser, emulsion, lipstick, makeup base, foundation, pressed powderand loose powder.
 11. A method of skin whitening or skin wrinklealleviation comprising applying the composition to claim 1 to skin. 12.A method of skin whitening or skin wrinkle alleviation comprisingapplying the composition to claim 5 to skin.